补肾养精颗粒调控早发性卵巢功能不全模型大鼠卵巢颗粒细胞凋亡机制研究  

Mechanism of Bushen Yangjing Granule(补肾养精颗粒)in Regulating Apoptosis of Ovarian Granulosa Cell in Model Rats with Premature Ovarian Insufficiency

作　　者：张芳[1] 丁怡[2] 于潇 刘鹏飞[1] 刘金星[1] ZHANG Fang;DING Yi;YU Xiao;LIU Pengfei;LIU Jinxing(Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,China)

机构地区：[1]山东中医药大学,山东济南230355 [2]山东中医药大学附属医院,山东济南250014

出　　处：《山东中医药大学学报》2022年第3期365-372,378,共9页Journal of Shandong University of Traditional Chinese Medicine

基　　金：山东省自然科学基金青年基金项目(编号:ZR2019PH053);山东省重点研发计划项目(编号:2019GSF108208)。

摘　　要：目的:探讨补肾养精颗粒抑制卵巢颗粒细胞凋亡治疗早发性卵巢功能不全的机制。方法:选择动情周期正常的SD雌性大鼠40只,随机分为空白组、模型组、中药组、激素组,每组各10只,模型诱导期间,空白组予生理盐水灌胃14 d,其他组予50 mg/kg雷公藤多苷片混悬液灌胃14 d建模。其后,空白组、模型组分别予生理盐水灌胃15 d,中药组予3.50 g/kg补肾养精颗粒灌胃15 d;激素组予0.1 mg/kg戊酸雌二醇片(第4天同时加用1 mg/kg醋酸甲羟孕酮片)灌胃,4 d为1个疗程,每4 d停用1 d,共15 d。通过免疫组化检测卵巢颗粒细胞中血管内皮生长因子(VEGF)、转化生长因子-β(TGF-β)、胰岛素样生长因子-1(IGF-1)及双向调控的核因子κB(NF-κB)、肿瘤坏死因子-α(TNF-α)表达;通过异硫氰酸荧光黄/碘化丙啶(Annexin-V-FITC/PI)荧光双染细胞凋亡及原位末端转移酶标记技术(TUNEL)检测卵巢颗粒细胞凋亡情况。结果:与空白组比较,模型组卵巢颗粒细胞VEGF、TGF-β、IGF-1水平显著下调,而NF-κB、TNF-α水平显著上调,细胞凋亡率显著增加(P<0.05);与模型组比较,中药组、激素组VEGF、TGF-β、IGF-1水平上调,NF-κB、TNF-α水平下调,细胞凋亡率下降(P<0.05);与激素组比较,中药组VEGF、IGF-1、TGF-β水平上调,NF-κB水平下调(P<0.05)。结论:补肾养精颗粒可通过上调卵巢颗粒细胞中VEGF、TGF-β、IGF-1的表达,降低NF-κB、TNF-α的表达,进而抑制卵巢颗粒细胞凋亡,促进卵泡发育。Objective:To explore the mechanism of Bushen Yangjing Granule(补肾养精颗粒)inhibiting ovarian granulosa cell apoptosis in the treatment of premature ovarian insufficiency.Methods:Forty female SD rats with normal estrous cycle were randomly divided into four groups including:blank group,model group,traditional Chinese medicine group and hormone group,ten rats in each group.During the model induction period,the blank group was intragastrically administered with normal saline for 14 days,and the other groups were intragastrically administered with 50 mg/kg tripterygium wilfordii polyglycoside tablet suspension for 14 days.After modeling,the blank group and model group were intragastrically administered with normal saline for 15 days,the traditional Chinese medicine group was intragastrically administered with 3.50 g/kg Bushen Yangjing Granule for 15 days.The hormone group was administered 0.1 mg/kg progynova(1 mg/kg medroxyprogesterone acetate tablets were added on the fourth day).Four days was a course of treatment.It was stopped for one day every four days,15 days in total.The expression of vascular endothelial growth factor(VEGF),transforming growth factor-β(TGF-β),insulin-like growth factor-1(IGF-1),bidirectional regulated nuclear factor kappa-B(NF-κB)and tumor necrosis factor-α(TNF-α)in ovarian granulosa cells was detected by immunohistochemistry.The apoptosis of ovarian granulosa cells was detected by Annexin-V-FITC/PI fluorescence double staining cell apoptosis and TUNEL.Results:Compared with the blank group,the levels of VEGF,TGF-βand IGF-1 of ovarian granulosa cells in the model group was significantly down regulated,while the levels of NF-κB and TNF-αwere significantly up-regulated and the apoptosis rate was significantly increased in the model group(P<0.05).Compared with the model group,the levels of VEGF,TGF-β,IGF-1 of ovarian granulosa cells in the traditional Chinese medicine group and hormone group was up-regulated,the levels of NF-κB、TNF-αwere down regulated,cell apoptosis rate wer

关 键 词：补肾养精颗粒 早发性卵巢功能不全 卵巢颗粒细胞 细胞凋亡 血管内皮生长因子 转化生长因子-Β 胰岛素样生长因子-1 核因子ΚB 肿瘤坏死因子-α 大鼠 

分 类 号：R285.5[医药卫生—中药学]